Background Diabetic retinopathy (DR) is a progressive vision-threatening complication of diabetes mellitus (DM),but its pathogenic mechanism is still unclear.Researches showed that growth arrest-specific gene product 6 (Gas6) /TAM system participates in pathogenesis and development of DR,and stromal-derived factors (SDF) vary in 2-type DM patients.However,whether Gas6/TAM and SDF-1 are associated with DR is below understood.Objective This study was to determine the relationship between the staging of DR and the levels of serum Gas6,SDF-1α and SDF-1β in DM patients.Methods A prospective cohord study was designed in this study.Ninety 2-type DM patients were included in the 3rd Affiliated Hospital of Southern Medical University Hospital from January to August in 2013.The patients were grouped into the non-diabetic retinopathy (NDR) group,background DR group (BDR) and proliferative DR (PDR) group,with 30 for each group.Thirty normal volunteers were enrolled in the same hospital and same period.The periphery blood 2 ml was collected from all the subjects under the consent inform.The levels of serum Gas6,SDF-1α and SDF-1β were assayed by ELISA,and leukocytes,neutrophils,plasma triglycerides (TG),total cholesterol (CHOL),high density lipoprotein cholesterol (H DL-C) and low density lipoprotein cholesterol (LDL-C) levels were detected and compared among the 4 groups.The correlations of serum Gas6,SDF-1 α and SDF-1 β changes with blood inflammatory cells and blood lipid were analyzed.Results The plasma CHOL concentrations were 4.93(4.14,5.44),5.02(4.35,5.69),4.54(3.85,5.93) and 5.99(5.11,6.89)mmol/L in the normal control group,NDR group,BDR group and PDR group,respectively,and the blood CHOL concentrations were significantly higher in the PDR group than those of the normal control group,NDR group,BDR group (P =0.002,P =0.007,P =0.006).White blood cell counts in the normal control group,BDR group were higher than those of the PDR group (P =0.034,P =0.015),neutrophil counts in the BDR group were higher than those of the PDR group (P =0.024),HDL-C in the NDR group was higher than that in the PDR group (P =0.032).LDL-C in the PDR group was higher than that in the normal control group.Compared with the normal control group,serum Gas6 levels were significantly lower in the NDR group and BDR group (P =0.048,P =0.006),and the serum Gas6 level showed an insignificant increase in the PDR group in comparison with BDR group (P =0.297).Serum SDF-1α levels in the PDR group was significantly higher than that in the BDR group (P =0.033) ;serum SDF-1β levels in the PDR group,BDR group were significantly higher than that in the NDR group (P =0.011,P =0.008) and normal control group (P =0.030,P =0.002).Weaker positive correlation was observed between the serum Gas6 and CHOL,TG,LDL-C levels (r=0.285,r=0.200,r=0.241,all at P＜0.05),between SDF-1α and SDF-1β (r=0.190,P＜0.05) as well as between SDF-1β and white blood cell (r=0.183,P＜0.05).Serum Gas6 served as dependent variable,while white blood cell,neutrophil,CHOL,TG,HDL-C,LDL-C,SDF-1α,SDF-1β served as independent variables,multiple stepwise regression analysis showed Gas6 =170.791 + 5.283CHOL (F =5.021,P =0.027).Conclusions Serum Gas6,SDF-1α and SDF-1β probably participate in the development of DR in 2-type diabetic patients.Gas6,SDF-1 α,SDF-1 β may play roles by affecting blood glucose level,angiogenesis,inflammatory cells and blood lipid metabolism.

To identify the active constituents in vitro and blood-absorbed ingredients in vivo from Yin Chen Hao decoction provides scientific evidence for probing its prevention and treatment mechanism on acute liver injury. An ultrahigh performance liquid chromatography quadrupole-time of flight-mass spectrometry (UPLC-QTOF/MS) method was applied for analysis of Yin Chen Hao decoction and the serum samples of mice with con-A induced acute liver injury after preventive oral administration for 14 days (the use of all laboratory animals in this study was approved by the Ethics Committee of the Naval Medical University, 19YF1459400). A total of 90 chemical constituents were identified from Yin Chen Hao decoction, mainly were flavonoids, terpenoids, tannins, quinones. 5 prototype compounds were identified in the serum, including chrysophanol, deoxyrhapontin-8-O-gallate, mussaenosidic acid, herniarin, emodin. The established UPLC-QTOF/MS method could efficiently and sensitively identify the constituents in vitro and blood-absorbed ingredients of Yin Chen Hao decoction, primarily clarify the material basis of its hepatoprotective effect, and provided a scientific basis for the quality marker selection and the pharmacodynamic material basis research on the decoction.

OBJECTIVE To determine the characterization, anti-tumor efficacy and pharmacokinetics of bufalin- loaded PEGylated liposomes compared with bufalin entity. METHODS Bufalin- loaded PEGylated liposomes and bufalin- loaded liposomes were prepared reproducibly with homogeneous particle size by the combination of thin film evaporation method and high pressure homogenization method. The particle size and zeta potential of the liposomes were determined by dynamic light scattering technique. The direct imaging of morphology of liposomes was charactered by transmission electron microscope. The content of bufalin in liposomes was analysed by HPLC method. The entrapment efficiency and the particle size was applied to assess the stability profile, after storage at 4℃ on day 0, 7, 15, 30 and 90. The in-vitro release behaviours of bufalin from liposomes were conducted using dialysis bag technique at 37℃. In-vitro cytotoxicity studies were carried out using MTT〔3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide〕assay on several kinds of tumor cell lines including SW620, PC-3, MDA-MB-231, A549, U251, U87 and HepG2. In-vivo pharmacokinetic study of bufalin liposomes was evaluated by HPLC method. RESULTS Their mean particle sizes were 127.6 nm and 155.0 nm, mean zeta potentials were 2.24 mV and - 18.5 mV, entrapment efficiencies were 76.31% and 78.40% , respectively. In- vitro release profile revealed that the release of bufalin in bufalin- loaded PEGylated liposomes was slower than that of bufalin-loaded liposomes. The cytotoxicity of blank liposomes has been found within acceptable range, whereas bufalin-loaded PEGylated liposomes showed enhanced cytotoxicity to U251 cells compared with bufalin entity. In-vivo pharmacokinetics indicated that bufalin-loaded PEGylated liposomes could extend eliminate half-life time of bufalin in plasma in rats. CONCLUSION The results suggested that bufalin-loaded PEGylated liposomes improved the solubility and increased the drug concentration in plasma.

We used 293 cells to express the recombinant membrane protein of the Ebola virus. Then, the immunogenicity of the recombinant protein was studied by immunized BALB/c mice. According to the codon use frequency of humans, the gene encoding the extracellular domain of the Ebola virus membrane protein was optimized, synthesized, and inserted into the eukaryotic expression plasmid pXG-Fc to construct the human IgG Fc and Ebola GP fusion protein expression plasmid pXG-modGP-Fc. To achieve expression, the fusion protein expression vector was transfected into high-density 293 cells using transient transfection technology. The recombinant protein was purified by protein A affinity chromatography. BALB/c mice were immunized with the purified fusion protein, and serum antibody titers evaluated by an indirect enzyme-linked immunosorbent assay (ELISA). Purification and analyses of the protein revealed that the eukaryotic expression vector could express the recombinant protein GP-Fc effectively, and that the recombinant protein in the supernatant of the cell culture was present as a dimer. After immunization with the purified recombinant protein, a high titer of antigen-specific IgG could be detected in the serum of immunized mice by indirect ELISA, showing that the recombinant protein had good immunogenicity. These data suggest that we obtained a recombinant protein with good immunogenicity. Our study is the basis for development of a vaccine against the Ebola virus and for screening of monoclonal antibodies.
Animals ; Antibodies, Viral ; immunology ; Ebolavirus ; genetics ; immunology ; Female ; Gene Expression ; Hemorrhagic Fever, Ebola ; immunology ; virology ; Humans ; Immunization ; Male ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; genetics ; immunology ; Viral Envelope Proteins ; genetics ; immunology

The epidemic of COVID-19 has brought great challenges to the global public health prevention and control system combined with clinical diagnosis and treatment system, and it makes the development of effective antiviral drugs an important task in current pharmaceutical research. Traditional Chinese medicine (TCM) has played an important role in the prevention and control of COVID-19. Due to its numerous chemical components and various structural types, TCM becomes a natural library for searching for lead compounds against SARS-CoV-2. In this study, a novel dual-target surface plasmon resonance (SPR) biosensor was developed for S protein receptor binding domain (SRBD) and angiotensin converting enzyme 2 (ACE2) which are two key proteins in the process of SARS-CoV-2 invading cells according to characteristics of synergistic effects of multiple components and comprehensive regulation of multiple targets of TCM. The SPR biosensor was applied to screen and identify active components from six TCMs, and daidzin from Puerariae Lobatae Radix was identified to bind with SRBD and ACE2. The affinity constant (K_D) of daidzin and ACE2 was 5.18 μmol·L^-1 through the SPR affinity assay. Competitive ELISA assay showed that daidzin could inhibit the binding of SRBD and ACE2, and the inhibition rate of daidzin (20 μmol·L^-1) was 38.6%. Molecular docking experiments further confirmed that daidzin had the best binding near the binding region of SRBD-ACE2 complex. This study shows that the dual-target SPR screening system is accurate and efficient, and is particularly suitable for screening of complex drug systems and effective substances study of TCM. It provides a material basis for exploring the mechanism of TCM active constituents against SARS-CoV-2, and provides a source of lead compounds for the development of anti-SARS-CoV-2 drugs.

The c-Abl nonreceptor tyrosine kinase is activated in the cellular responses to genotoxic, oxidative and other forms of stress. Using tagged forms of c-Abl, the present studies demonstrate that c-Abl forms homodimers in cells. The results show that the c-Abl N-terminal regions interact with the corresponding C-terminal regions of both partners in the dimmer. Specifically, the c-Abl SH3 domain binds to a proline-rich motif at amino acids 958-982 in the c-Abl C-terminal region. Deletion of the proline-rich motif disrupts dimmer formation. These findings provide the first evidence that c-Abl forms homodimers and indicate that homodimerization can contribute to the regulation of c-Abl activity.
Humans ; Protein Multimerization ; Proto-Oncogene Proteins c-abl ; genetics ; metabolism ; src Homology Domains

Malignant proliferating tricholemmoma is a very rare dermatic annexal tumor originated from outer root sheath cells. In this article, a case of facial multiple malignant proliferating tricholemmoma was reported, and its clinical pathologic features, differential diagnosis, treatment methods and histogenesis were discussed by reviewing relevant literatures.
Diagnosis, Differential ; Face ; Female ; Humans ; Skin Neoplasms

XYL1 gene,which encodes xylose reductase with dual coenzyme activity from Candida tropicalis,was transformed into Pichia pastoris X-33 by expression vector pGAPZB.The recombination strain was immobilized in Ca-alginate beads and fermentation characterization is studied using corn cob hydrolysates.Fermentation conditions were as follow:initial pH value 6.0,30℃,initial cell concentration of 20%,the Liquid volume of 28%,rotation speed 130r/min.The average xylitol yield was 37.5% on the optimum condition.This result is expected to provide a new alternative method for producing xylitol on a large scale by bioconversion.

OBJECTIVE: To explore the effect of hepatocyte growth factor (HGF) on oxygen-glucose deprived injury and apoptosis of astrocytes. METHODS: The injury of primary cultured rat cerebral cortical astrocytes was induced by oxygen-glucose deprivation. Astrocytes were treated with HGF at various final concentrations of 20 - 100 ng/mL. The cell damage and viability were evaluated by the lactate dehydrogenase (LDH) released rate and the 3- (4,5-dimethylthazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) conversion method. Detection of apoptotic cells was determined by the flow cytometry, and the ultrastructure was observed by the transmission electron microscope. RESULTS: Oxygen-glucose deprivation increased the LDH release rate, decreased the cell viability and increased the number of apoptotic astrocytes. While exposed to HGF at the same condition, the LDH release rate decreased, the cell viability increased, and the percentage of apoptotic cells decreased (P <0.05). The maximum protective effect of HGF was observed at 60 ng/mL. CONCLUSION HGF can protect cultured astrocytes from oxygen-glucose deprived injury, and attenuate the apoptosis of astrocytes in a dose-dependent manner.
Animals ; Apoptosis ; drug effects ; Astrocytes ; pathology ; Cell Hypoxia ; Glucose ; pharmacology ; Hepatocyte Growth Factor ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism

Objective Aselectivitystudywasconductedthroughtheexaminationofradionuclides137Cs(Cesium-137)for foodbasedondifferentdetectionconditions.Methods Atotalof48foodsampleswereselectedfromthreeareasincluding Qinshan nuclear power plant,Sanmen nuclear power plant and Hangzhou and Zhoushan respectively.1 37 Cs of these samples were determined by γspectrometry and Phosphoric acid ammonium molybdate method.The level of 48 foods were statistically analyzed,and then the time consuming,sample size requirements,influence factors were comprehensively discussed,thustheselectionreferenceproposaloftheexaminationmethodcouldbeprovided.Results Therewereno significant difference for the data of two examination method (P>0.05 ).The limit of detection of the γspectrometry was lower (P <0.05 ).Compared with Phosphoric acid ammonium molybdate method,γspectrometry had lower limit of detection,and could detect a variety of radionuclides at a time,but need more sample and time-consuming when multi-sample were detected.The limit of detection of the Phosphoric acid ammonium molybdate method was high,and the chemicalprocessingstepswerecumbersomeandwaseasytobeinterferedby134Cs.Conclusion Thelimitofdetectionof the γspectrometry is low,and the sensitivity of the Phosphoric acid ammonium molybdate method is high.Most food are recommended to be detected by γspectrometry in the practical work,and the food which were difficult collected,less ash or low content,are recommended to be detected by Phosphoric acid ammonium molybdate method.